ES2328822T3 - OLIGO-BETA- (1,3) -GLUCANO AND MONOCLONAL ANTIBODIES AGAINST CANCER. - Google Patents

Abstract

The present invention relates to compositions and methods for treating humans and warm-blood animals suffering from cancer. More particularly, a therapeutical treatment in which a monoclonal antibody is administered with either beta-(1,3)-glucan like laminarin or a oligo-beta-(1,3)-glucan and a pharmaceutically acceptable carrier, to patients suffering from cancer are described.

Description

Oligo- ?- (1,3) -glucan and monoclonal antibodies against cancer.

Technical sector

The present invention relates to a therapeutic treatment in which an antibody is administered monoclonal with a oligo- ?- (1,3) -glucan a patients suffering from cancer and drug use in said treatment.

More particularly, it refers to a method of treatment of humans and warm-blooded animals that They have cancer.

State of the art

Glucans that have been thoroughly studied they are natural products, and are known for presenting activities of immunostimulation However, it has been observed that not all Compounds comprised in glucans that are produced in a manner Natural are active.

Among the glucans already studied, it can be mentioned laminarin for presenting immunostimulatory activities and for consequently be useful in therapeutic treatments, in particular for a patient suffering from cancer, as disclosed, by example, in international patent application WO03 / 045414 a Name of the present inventors.

In addition, applicants have also discovered  that some of the oligo- ?- (1,3) -glucans specific also have an immunostimulation activity. Those oligo- ?- (1,3) -glucans They are the ones that present the following formula:

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one

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in which n = from 1 to 10, preferably, n = 2 or n = 3, or a pharmaceutically salt acceptable of same.

Monoclonal antibodies are prepared normally fusing an antibody producing B cell, normally short-lived, (see immunity) with a cell of rapid growth, such as a cancer cell (sometimes called "immortal" cell). The resulting hybrid cell, or hybridoma, multiplies rapidly, creating a clone that produces large amounts of the antibody. Monoclonal antibodies engendered much enthusiasm in the world of Medicine in the years 1980, especially as possible cures for cancer.

In order to use these antibodies in cancer treatments, researchers need find antigens on the surface of cancer cells that found only in those cancer cells and that is not found in normal tissues, and then produce antibodies monoclonal against those antigens.

The theory is that these monoclonal antibodies they could then recognize the antigen in cancer cells and attach to it (like a key in a lock). This could then trigger that the body's immune system attack on cancer cells. Alternatively, the antibody monoclonal could have a cancer drug or a radioactive substance bound thereto and used to deliver the treatment directly to cancer (this was called therapy directed or the "panacea").

In the last twenty-five years they have addressed a lot of research both in the search for antigens in cancer cells as in the improvement of production of monoclonal antibodies so that the amounts necessary for medical use.

However, the demand for Cancer treatments that are effective and not harmful.

Miyanishi et al . (J of Bioscience and Bioengineering, vol. 95, No. 2, 2003, pp. 192-195) teaches the use of a human anti-TNF? Polyclonal goat antibody or the use of goat anti-TNF polyclonal antibody Human β in determining whether the cytotoxic activity of MC-CM prepared with an oligosaccharide is affected by said antibody.

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In WO 98/39013 A, it is disclosed a pharmaceutical product for the treatment of tumors in beings human comprising β- (1,3) -glucan and anti-tumor antibody Neither of these two documents discloses the synergistic combination of this patent request.

In WO02 / 058711, the use of laminarin in combination with the monoclonal antibody, but glucans that have a higher molecular weight are considered, In particular, barley glucans are the most effective.

Similarly, EP 0 194 851 teaches a pharmaceutical product for the treatment of tumors in human beings comprising the β- (1-3) -glucan lentinano with monoclonal antibodies of the type IgG2a and IgG3.

Surprisingly and unexpectedly, despite the teachings of document W002 / 058711, the present inventors discovered that the monoclonal antibody and the oligo-? - (1,3) -glucan have a synergistic effect in the treatment of cancer.

Object of the invention

The present invention is based on said effect synergistic

Therefore, an object of the present invention is  a therapeutic method comprising the administration of a composition comprising a monoclonal antibody with a oligo- ?- (1,3) -glucan and a pharmaceutically acceptable vehicle, to a human being or to a warm-blooded animal suffering from cancer in an amount that is Effective to treat cancer.

Another object of the invention is the use of a composition comprising a monoclonal antibody with a oligo- ?- (1,3) -glucan and a pharmaceutically acceptable vehicle, for the manufacture of a medicine for the treatment of cancer.

Throughout the specification, the amount of active composition is considered "effective" if allows obtaining the intended medical purpose such as control or destruction of cancer cells without producing unacceptable toxic symptoms. That effective amount will vary with factors such as the particular state being treated, the physical condition of the patients and the duration of the treatment.

The "acceptable pharmaceutical vehicle" is select from the group comprising vehicles, agents of suspension or pharmaceutically acceptable solvents, and depending of the chosen route selected for administration, and having take into account conventional pharmaceutical practice; "acceptable" means that the vehicle is compatible with the other components of the formulation and is not harmful to the patient.

More generally, a "component pharmaceutically acceptable "must not present or induce too many adverse side effects such as toxicity, irritation and an allergic response, and should be consistent with a reasonable benefit / risk ratio.

He oligo- ?- (1,3) -glucan is a compound having the following formula (1):

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2

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in which n = from 1 to 10, preferably, n = 2 or 3, or a pharmaceutically acceptable salt of the same.

Advantageously, the oligo- ?- (1,3) -glucans assets are those of formula I above,

in which n = 2, that is, β-D-glucopyranosyl- (1 \ rightarrow 3) -? -D-glucopyranosyl- (1 \ rightarrow 3) -? -D-glucopyranosyl- (1 \ rightarrow 3) - β-D-glucopyranoside, which is called laminaritetraose, or in which n = 3, that is the β-D-glucopyranosyl- (1 \ rightarrow 3) -? -D-glucopyranosyl- (1 \ rightarrow 3) -? -D-glucopyranosyl- (1 \ rightarrow 3) -? -D-glucopyranosyl- (1 \ rightarrow 3) - β-D-glucopyranoside, which is called laminaripentaose.

These compounds can be synthesized by deprotection and purification of the compounds prepared according to the procedure disclosed in document WO01 / 57053 in the name of the assignee. Deprotection and purification methods that can used are described by reference to laminaribious in the document FR2777281.

In the method according to the invention, the antibody monoclonal is any specific monoclonal antibody against molecular determinants present in cancer cells and that You can simultaneously activate the plug-in.

Advantageously, the monoclonal antibody is  select from the group comprising Herceptin, alemtuzumab, rituximab, tositumomab, Campath, cetuximab (Erbitux (D), edrecolomab, Mylotarg, Panorex, pentumomab.

The method according to the invention is suitable for treat cancer, specifically, leukemia, adenocarcinoma, cancer breast, lung cancer, ovarian cancer, esophageal cancer, stomach cancer, intestinal cancer, non-Hodgkin lymphoma or cancer of colon

Since the monoclonal antibody is manage with a oligo- ?- (1,3) -glucan, the Treatment may be called "combination therapy."

Combination therapy can be sequential, which means that the treatment is carried out first with a agent and then with the second agent; or it can be simultaneous, what which means that both agents are administered at the same time.

In the method according to the invention, the amount effective or of β- (1,3) -glucan as laminarin or oligo- ?- (1,3) -glucan is 2 at 20 mg / kg when administered orally for treatment sequential. The amount of monoclonal antibody is the amount Conventional used in the treatment of cancers.

According to another object of the invention, the method according to the invention further comprises the administration of an agent Chemotherapeutic for enhanced potentiation.

The method according to the invention can be used. also in combination with radiotherapy treatment.

According to another object of the invention, when the method further comprises the administration of an agent chemotherapy or a radiotherapy phase treatment, the monoclonal antibody can be administered in combination with laminarin

Laminarin is a glucan that is produced from natural way that is extracted from brown algae and that consists of polysaccharides with an average molecular weight between approximately 2,500 and 6,000.

The assignee markets a laminarin commercially available for other purposes.

The present invention also relates to compositions useful in the mentioned therapeutic method previously.

According to the present invention, the composition for its use in a sequential or simultaneous treatment can be administered intravenously to the patient, in the form of injections, ointment, pulmonary spray; for use in a sequential treatment, the Composition can also be administered as follows: monoclonal antibody is administered intraperitoneally or by IV, while the oligo- ?- (1,3) -glucan, or possibly laminarin, is administered orally to the patient, in the form of a solution, a suspension, a syrup, a tablet, one capsule

It can also be presented as an injection in Bolus, an electuary or a paste.

The oral formulations of oligo- ?- (1,3) -glucan, or possibly laminarin, suitable for sequential treatment in relation to the practice of the present invention include capsules, gels, seals, tablets, granules, tablets or powders  effervescent or non-effervescent; they can consist of a solution, or suspension in an aqueous or non-aqueous liquid, in a liquid oil emulsion in water or in a water emulsion in oil.

Generally, said oral formulations can  be prepared by uniformly mixing the active substance, that is, especially the oligo-? - (1,3) -glucan soluble, or possibly laminarin, possibly together with an agent chemotherapeutic, with liquid vehicles or solid vehicles finely divided or both, and then, if necessary, shaping the product

Suitable solid vehicles comprise lactose, sucrose, gelatin, agar agar and bulk powders.

Suitable liquid vehicles include water, pharmaceutically acceptable fats and oils, alcohols or others organic solvents, including esters, emulsions, syrups or elixirs, suspensions, solutions and / or suspensions, and reconstituted solutions and / or suspensions from granules non-effervescent and effervescent preparations reconstituted to from effervescent granules.

They may also contain, for example, preservatives, emulsifying agents, suspending agents, diluents, sweeteners, thickeners and melting agents; vehicles Preferred liquids are edible oils, for example, oils of corn or canola, as well as polyethylene glycols (PEG).

Therapeutic forms, intended for oral administration, may comprise an inert vehicle, not toxic, pharmaceutically acceptable, selected from the group that It comprises lactose, starch, sucrose, glucose, methylcellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol, cyclodextrin and cyclodextrin derivatives.

Capsules or tablets containing a oligo- ?- (1,3) -glucan, or possibly laminarin, according to the invention should be formulated preferably simple and easy to swallow or chew. The tablets may contain vehicles, binders, lubricants, diluents, disintegrating agents, coloring agents, flavoring agents, flow induction agents or adequate fusion. A tablet can be produced by compression or molding, optionally with one or more components Additional classics

Tablets manufactured by compression compressing the active substance in a loose form (e.g. powders, granules) optionally mixed with a binder (for example, gelatin, hydroxypropyl methylcellulose), lubricant, inert diluent, preservative, disintegrant (for example, starch sodium glycolate, gelatin, natural sugars such like glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as sodium alginate, carboxymethyl cellulose, polyethylene glycol, waxes. Lubricants used in these pharmaceutical forms include sodium oletao, sodium stearate, magnesium stearate, sodium acetate, sodium chloride. Disintegrating agents include, for example, starch, methylcellulose, agar agar, bentonite, xanthan gum. The molded tablets are prepared by molding in a suitable machine  a mixture of the powdered active ingredient moistened with a inert liquid diluent.

The tablets are optionally coated and can be formulated so as to provide a slow release or controlled of the active substance. The tablets may be also optionally equipped with an enteric coating for provide a release in parts of the intestine other than stomach.

Detailed description of the invention

The following examples are intended for illustrate the invention, in particular, to illustrate the synergistic effect of a monoclonal antibody, specifically Herceptin, with laminarin on tumor growth.

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Examples

In the examples the following are used products:

Phycarine®:

laminarin extracted from brown algae by Laboratoires Göemar.

Herceptin:

manufactured by Genentech,

laminaritetraose:

synthesized according to the document US10 / 668,665,

laminaripentaose:

synthesized according to the document US10 / 668,665.

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Example 1 Effect on tumor growth

Naked female mice ( nu / nu ) between 5 and 6 weeks of age weighing approximately 20 g were obtained from Harlan, Inc. (Madison, WI). The human breast carcinoma cell line BT-474 from the American Type Culture Collection (ATCC) was obtained. BT-474 was established by E. Lasfargues and WG Coutinho from a solid breast carcinoma obtained from a 60-year-old female patient (2). Twenty-one-day 17β-estradiol microgranules were implanted at 0.25 mg (Innovative Research of America) subcutaneously in each mouse. The next day, the animals were implanted sc through trocar fragments of human tumor carcinomas collected from sc growing tumors in host mice. When the tumors were approximately 40 mg in size (24 days after inoculation), the animals were equally divided into treatment and control groups. All groups contained 9 mice. All mice were tagged in the ear and their individual follow-up was carried out throughout the experiment.

Mice from each group were given  intraperitoneally 2 ml of the following composition:

Group 1:

(control) Sterile H2O, 5 times at day

Group 2:

Herceptin 0.5 mg / kg, twice a week for 3 weeks

Group number 3:

Phycarine® 250 mg / kg, once daily for 5 days

Group 4:

Phycarine® 250 mg / kg, once daily for 5 days Herceptin 0.5 mg / kg, twice a week for 3 weeks

Team 5:

Phycarine® 500 mg / kg, once daily for 5 days Herceptin 0.5 mg / kg, twice a week for 3 weeks

Group 6:

(reference) Paclitaxel (Mead Johnson) 16 mg / kg, once a day for 5 days.

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The mice were weighed twice a week, and tumor measurements were obtained using calibrators twice a the week, starting on day 1. These tumor measurements are converted to mg of tumor weight by the conventional formula, (W 2 x L) / 2 (2). The experiment is over when the size Tumor of the control group reached an average of 500 mg. Behind the completion (day 46), mice were weighed, sacrificed and  They removed their tumors. The tumors were weighed and the average tumor weight per group. In this model, the change in weight Tumor treated medium / change in tumor weight control medium x 100 was subtracted from 100% giving tumor growth inhibition (TGI) for each group.

Paclitaxel produced tumor remission in this tumor xenograft model. With this agent, the weight was subtracted end of a given tumor of its own weight at the start of treatment on day 1. This difference was divided by the initial tumor weight giving the% of remission. An average% of tumor remission was calculated at from data from mice in a group that experienced remissions Tumor

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(Table goes to page next)

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The results that are expressed as the value mean (standard deviation) are given in the following table one.

TABLE 1

3

Graphical representation was performed using GraphPad Prism® software and is provided in Figure 1, in the that, for each group, the corresponding graph represents the weight of the tumor (in mg) depending on the day (from 1 to 46).

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These results show that:

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the administration of Phycarin® 500 mg / kg does not allow a limitation of tumor growth, but quite the opposite, enhances the tumor growth;

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the administration of Herceptin 0.5 mg / kg allows a limitation of tumor growth;

-

the administration of rnycarin) 250 mg / kg allows a limitation of tumor growth that is approximately the same as the obtained by administering Herceptin 0.5 mg / kg;

-

the administration of Phycarin® 500 mg / kg and Herceptin 0.5 mg / kg allows a limitation in the increase in tumor weight that is much greater than the average value obtained when administering Herceptin 0.5 mg / kg and Phycarin® 500 mg / kg alone; said activity being on the tumor weight even equivalent to that obtained when administered a conventional dosage of taxol.

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Example 2

Female nude nude mice ( nu / nu ) between 4 and 6 weeks of age weighing approximately 20 g were obtained from Jackson Laboratories, USA. The human breast carcinoma cell line BT-474, obtained from the American Type Culture Collection (ATCC), was injected into 0.1 ml of PBS in the mammary glands of each mouse. All groups contained 6 mice. All the animals were tagged on the ear and their individual follow-up was carried out throughout the experiment.

When the tumor diameter reached 0.7 to 0.9 mm, usually 14 days after the injection of tumor cells, The treatment was started.

Mice were administered daily for two weeks the following:

Group 1:

(control) sterile PBS, 5 times a day

Group 2:

laminaritetraose (i.p.) 100 \ mug / mouse,

Group number 3:

laminaritetraosa (i.p.) 100 \ mug / mouse more Herceptin (IV) 0.5 mg / kg,

Group 4:

laminaripentaose (i.p.) 100 \ mug / mouse,

Team 5:

laminaripentaose (i.p.) 100 \ mug / mouse more Herceptin (IV) 0.5 mg / kg.

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The mice were weighed, two were sacrificed weeks after the start of treatment, the tumors, the surrounding tissue was trimmed and weighed.

The results that are expressed as the value mean in mg are represented in figure 2. In said figure, the mean tumor weight for each group of mice is indicated in the ordered as mg.

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These results show that:

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the administration of either laminaritetraose or of laminaripentaous allows a decrease in the weight of tumor;

-

the administration of either laminaritetraose or of laminaripentaosa and Herceptin allows a decrease in the weight of the tumor that is much greater than the average value obtained when administer laminaritetraose or laminaripentaose alone.

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References cited in memory

This list of references cited by the applicant is directed only to help the reader and not form part of the European patent document. Even if you have tried the greatest care in its conception, errors cannot be excluded or omissions and the OEB declines all responsibility to this respect.

Patent documents mentioned in the report

WO 03045414 A (0004)

WO 0157053 A (0024)

WO 9839013 A (0012)

FR 2777281 (0024)

WO 02058711 A (0013) (0015)

US10668665 B (0049) (0049)

EP 0194851 A (0014)

Non-patented literature cited in the description

\ bullet Miyanishi et al. J. of Bioscience and Bioengineering 2003 , vol. 95 (2), 192-195 (0011).

Claims (15)

1. Product containing:

-

a specific monoclonal antibody against molecular determinants present in cancer cells and that simultaneously can activate the add-on, and

-

a oligo-? - (1,3) -glucan from Formula 1)

5 in which n = from 1 to 10, preferably n = 2 or 3, or an acceptable pharmaceutical salt of the same, as combined preparation for simultaneous, separate use or sequential in a method to treat cancer in humans or in a blood animal hot. 2. Product according to claim 1, in the that the monoclonal antibody and the oligo- ?- (1,3) -glucan is administered intravenously or intraperitoneally patient in the form of injections, or in the form of ointments or pulmonary sprays 3. Product according to claim 1, in the that

-

he monoclonal antibody is administered intraperitoneally or by intravenous route, and

-

he oligo- ?- (1,3) -glucan is administered orally to the patient in the form of a solution, a suspension, a syrup, a tablet or a capsule.

4. Product according to claim 3, wherein the oligo-? - (1,3) -glucan is administered in an amount of 2 to
20 mg / kg 5. Product according to any one of the claims 1-4, wherein the cancer is leukemia, adenocarcinoma, breast cancer, lung cancer, cancer of ovaries, esophageal cancer, gastric cancer, intestinal cancer, non-Hodgkin lymphoma or colon cancer. 6. Pharmaceutical composition comprising:

-

a specific monoclonal antibody against molecular determinants present in cancer cells and that simultaneously can activate the addon,

-

a oligo-? - (1,3) -glucan from Formula 1)

6

-

in the that n = 1 to 10, preferably n = 2 or 3, or a salt acceptable pharmaceutical thereof, and

-

a pharmaceutically acceptable vehicle, for use in a method for treat cancer in humans or in a blood animal hot.

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7. Pharmaceutical composition according to claim 6, wherein the cancer is leukemia, adenocarcinoma, breast cancer, lung cancer, ovarian cancer, cancer esophagus, gastric cancer, intestinal cancer, non-Hodgkin lymphoma or colon cancer. 8. Pharmaceutical composition according to claim 6 or 7, wherein said method for treating cancer is characterized in that the monoclonal antibody and oligo-? - (1,3) -glucan are administered simultaneously, sequentially or successively. 9. Pharmaceutical composition according to claim 8, wherein the monoclonal antibody and the oligo- (3-? - (1,3) -glucan is administered intravenously or intraperitoneally patient in the form of injections, or in the form of ointments or pulmonary sprays; or are administered as follows: monoclonal antibody is administered intravenously while that oligo- ?- (1,3) -glucan it is administered orally to the patient, in the form of a solution, a suspension, a syrup, a tablet, a capsule. 10. Pharmaceutical composition according to a any one of claims 6 to 9, wherein said method to treat cancer includes oral administration from 2 to 20 mg / kg of oligo-? - (1,3) -glucan. 11. Pharmaceutical composition comprising:

-

a specific monoclonal antibody against molecular determinants present in cancer cells and that simultaneously can activate the addon,

-

a oligo-? - (1,3) -glucan from Formula 1)

7

in which n = of 1 to 10, preferably n = 2 or n = 3, or a pharmaceutically salt acceptable thereof, and

-

a acceptable pharmaceutical carrier, said composition being free of any other glucan.

12. Pharmaceutical composition according to claim 11, said composition being in the form of a injection, an ointment or a pulmonary spray. 13. Pharmaceutical composition according to claim 11 or 12, further comprising an agent chemotherapeutic 14. Pharmaceutical composition comprising:

-

a specific monoclonal antibody against molecular determinants present in cancer cells and that simultaneously can activate the addon,

-

a oligo-? - (1,3) -glucan from Formula 1)

8

in which n = of 1 to 10, preferably n = 2 or n = 3, or a pharmaceutically salt acceptable thereof, or laminarin,

-

a chemotherapeutic agent, and

-

a pharmaceutically acceptable vehicle, for use in a method for treat cancer in humans or in a blood animal hot.

15. Pharmaceutical composition comprising

-

a specific monoclonal antibody against molecular determinants present in cancer cells and that simultaneously can activate the addon,

-

a oligo-? - (1,3) -glucan from Formula 1)

9

in which n = of 1 to 10, preferably n = 2 or n = 3, or a pharmaceutically salt acceptable thereof, or laminarin, and

-

a pharmaceutically acceptable vehicle, for use in a method for treat cancer in humans or in a blood animal hot, said method further comprising a phase treatment of radiotherapy